EA043403B1 - PELLET WITH MULTILAYER STRUCTURE FOR SLOW RELEASE OF ACTIVE INGREDIENT INTO THE DISTAL SECTION OF THE COLON - Google Patents

Description

For the treatment of inflammatory processes and changes in the distal parts of the colon, for example ulcerative colitis, a dosage form is needed that, after oral administration, ensures local availability of the active substance, which is budesonide, in a sufficiently high concentration at the site of inflammation. This concept, also called colon targeting, is not feasible using a traditional oral pharmaceutical composition because there is a high risk that the active substance cannot be present in effective concentrations at the site of action, i.e. in pathologically altered parts of the colon. Therefore, the problem of targeting the colon should preferably be solved by using a composition with a modified release of the active substance, which is released as completely as possible in the colon and can exert its effect there. The present invention is particularly suitable for the preparation of a dosage form that is simple and reliable in administration and satisfies these requirements.

The prior art describes a wide variety of compositions for the treatment of inflammatory bowel diseases. There are different manifestations of inflammatory bowel disease, and Crohn's disease can be found in different parts of the intestine. In some forms of inflammatory diseases, the focus of inflammation is in the duodenum. In other forms of manifestation of inflammatory diseases, the focus of inflammation is in the jejunum or ileum or in the colon. In some patients, the affected areas extensively involve many parts of the intestine. Ulcerative colitis manifests itself almost exclusively in the large intestine, which is also divided into different sections. In most cases, the rectum is affected, from where the disease spreads to the sigmoid colon and colon.

In pharmaceutical technology, there is a problem in ensuring the presence of the active substance as precisely as possible in the place of the intestine where there is a need for the active substance. This task of ensuring the release of the active substance preferentially in the colon is not satisfactorily solved by compositions known from the prior art.

WO 91/07172 describes orally administered compositions for the treatment of inflammatory bowel diseases, one of the active ingredients being budesonide. The pellets described in this publication have a structure in which a layer of active substance is applied to the original cores, which are coated with two different layers, which are sprayed onto carrier pellets with a layer of active substance. WO 95/08323 describes pellets containing budesonide with a controlled release pattern of this active substance. Such pellets contain, from the inside to the outside, neutral pellets, a layer of active substance, an inner varnish layer of varnishes soluble in intestinal juice and an outer varnish layer of varnishes insoluble in gastric juice, soluble in intestinal juice. The post-gastric release profile achieved in this way cannot give satisfactory results for the indication described above (ulcerative colitis). The required slow, continuous release of the active substance in the colon is not achieved. Hard capsules which contain such pellets as described in the prior art are characterized by an early, immediate and rapid release of the active substance, which for this reason generally reaches the colon only in very small quantities.

Pharmaceutical compositions with multilayer coatings are also described in WO 03/045356 or WO 2017/216088, where mucoadhesive materials are also provided. Further compositions for controlled release of the active substance are described in WO 2009/138716 or WO 00/76478, wherein the active substances are partially embedded in three-dimensional matrices, thereby achieving a sustained release. Complex medicinal compositions that continuously release the active substance from the matrix are also known (cortiment-MMX delayed-release tablets). WO 02/17887 describes medicinal products for the treatment of intestinal diseases, wherein the active substance is released mainly in the distal parts of the intestine. These are pellets, granulates or mini-tablets, which predominantly contain 5-aminosalicylic acid and are individually coated with a stomach acid-resistant varnish and a second varnish.

EP 2143424 describes dosage forms for use specifically in the colon. The composition contains a core coated with a layer of active substance. On top of this layer is applied an intermediate layer of a cationic polymer, which is capable of swelling at a pH value of no more than 6.6, and an outer layer, which is an anionic polymer that is soluble at a pH value of no less than 7.0. The release of the active substance occurs after a delay of approximately 300 minutes relatively quickly, abruptly and completely.

DE 4332394 describes budesonide-containing controlled-release pellets, wherein the neutral pellets have a layer of budesonide as active substance and excipients and two different varnish layers.

WO 03/080032 describes pharmaceutical compositions which contain essentially a core with budesonide as active substance, a middle layer of coating of soluble intestinal

- 1 043403 with a polymer composition and an outer coating resistant to the action of gastric juice.

EP 1607087 describes oral compositions for use in the colon.

In Gross et al. in Journal of Crohn's and Colitis, 5, 2011, p. 129-138, reports a study in which it was found that treatment with mesalazine granules at a dose of 3 g (Salofalk®) leads to better results than treatment with oral budesonide at a dose of 9 mg, since when using known in the art containing budesonide preparations could not reliably ensure the release of the active substance in the colon.

A common feature of the pharmaceutical compositions known from the prior art is that the required release of budesonide as active substance is not ensured in such a way that this active substance is almost completely present in the inflamed areas throughout the colon and especially also in the rectum.

The subject of the present invention is multi-layer coated pellets for delayed and continuous release of the active substance throughout the colon (Fig. 1), the pellet having the following components (Fig. 2):

a) the original pellet, which consists only of inert material and does not contain any pharmaceutically active active substance inside;

b) a layer with the active substance, which is applied directly to the original pellet and, in addition to the active substance, contains only excipients commonly used in pharmaceuticals;

c) a swelling layer, which is applied directly to the active substance layer and contains swelling materials that swell upon contact with intestinal juice;

d) a sustained release layer, which is also insoluble in intestinal juice at a pH value greater than 6.5, but becomes permeable to liquids, and which is applied directly to the swelling layer, as well as applied directly to it;

e) an outer coating that does not dissolve at a pH value less than 5.5, but readily dissolves at a pH value greater than 6.0.

In one preferred embodiment, no other intermediate layers are located between the individual layers mentioned above. This means that layer (b) with the active substance is applied directly to the original pellet (a), and a swelling layer (c) is applied directly to the layer with the active substance. The swelling layer is directly coated with a coating (d), which is a sustained release layer, and directly coated with an outer coating (e), which is resistant to gastric acid.

The present invention provides a pharmaceutical composition which contains budesonide or a pharmaceutically acceptable salt thereof as an active ingredient and which is preferably used for the treatment of inflammatory diseases of the colon.

The original pellet(s) used have an average diameter of 0.2 to 2.0 mm, with at least 90% of the particles having sizes within these limits. In addition, the original pellets have a spherical shape with a uniform surface.

Pharmaceutical pellets are approximately spherical particles with an average diameter of 0.2 to 2 mm, with a uniform surface and a narrow size distribution. The surface of such particles is smooth and low-porous.

It is important that the balls used as starting pellets meet certain quality requirements. The pellet material used is sugar pellets, which consist of sucrose and corn starch, and the sucrose content should not exceed 90 wt.%. Pellets preferably have a narrow size distribution, since this is one of the prerequisites for a uniform distribution of the finished pellets. The particles have a diameter of 0.2 to 2.0 mm, preferably 0.7 to 1.4 mm, with a particularly preferred range of 0.85 to 1.0 mm. When producing pellets, it should be taken into account that their size distribution varies within relatively narrow limits, and therefore, for example, at least 90%, preferably at least 95%, of the particles have sizes within the specified limits. Methods for determining particle size and determining their size distribution are described in the European Pharmacopoeia. In this regard, reference is made to Chapter 2.9.38 of this pharmacopoeia.

Another important aspect concerns the nature of the pellet surface. The original pellets should correspond to the maximum possible extent to the shape of an ideal ball, and their surface should be as smooth as possible without depressions or external protrusions. The quality of pellets can be monitored, for example, using a stereo microscope and a digital (filming) camera attached to it. The resulting images can be processed using software suitable for these purposes.

According to the invention, budesonide, which is a local corticosteroid, is used as the active substance. Budesonide is practically insoluble in water. Therefore, in

- 2 043403 Preferably, micronized budesonide is used.

For medical use, the use of micronized budesonide is particularly suitable (particle size distribution specification: 100% of particles have a size less than 10 μm and at least 95% of particles have a size less than 5 μm, determination method: laser diffraction). The physicochemical properties of budesonide are known (literature: for example, Merck Index reference book, commentary on the pharmacopoeia):

crystalline powder with a color ranging from white to almost white;

solubility in aqueous systems practically does not depend on the pH value and is 0.014 mg/ml; the specific rotation [a] ^D 2 ₀ of epimer A is +98.9 (0.28%, in dichloromethane);

pKa value: 12.85±0.10;

melting point: 221-232°C.

When choosing the form of administration proposed in the invention, it is assumed that the continuous release of the active substance occurs from the intact dosage form and that the duration of its presence in the gastrointestinal tract and the duration of its passage through it do not have an adverse effect (for example, due to a longer stay in the stomach after eating). The dose of the medically active component can be distributed over an almost uniform, but at the same time large outer surface and flexibly adjusted. Such properties are achieved with the multi-particle application form proposed in the invention, namely pellets. Packing pellets in a certain quantity into hard capsules or portioned packaging, such as stick packs, provides the possibility of regulating, taking and using budesonide in a strictly defined dose. The properties of the medicinal composition described in a particularly preferred embodiment make it possible to administer budesonide into the body in a single dose varying from 3 to 9 mg.

The use of a medicinal composition containing budesonide proposed in the invention significantly reduces the risk of unwanted absorption of budesonide into the systemic circulation. The multiparticulate form of application has the advantage over the one-piece compositions described in the prior art that it provides reproducible and reliable, i.e. less subject to fluctuations, release of the active substance over a large area of the inflamed colon, and is therefore particularly suitable for the treatment of parts of the intestine affected by inflammation. Thus, the dose of budesonide used can be administered into the body distributed over a large surface area, which is located in the area of the colon.

Premature release of the medically active component from the application form according to the invention in the stomach is prevented due to the resistance of the outer coating (e) to the action of gastric juice. Thanks to this gastric juice-resistant outer coating (e), the outer shell of the pellets does not dissolve in the stomach. In the stomach, the prevailing pH value is from about 1 to a maximum of about 5. While the pellets are in the stomach area, the outer shell layer does not dissolve. Only when passing into the small intestine does the pH value increase and the outer coating (e) dissolves. In the small intestine and depending on time, intestinal juice penetrates through the second outer shell layer (d) into the pellets. The sustained release layer (d) does not dissolve upon contact with intestinal juices. However, due to the permeability of this layer to intestinal juice, it comes into contact with the underlying swelling layer (c). As a result of contact with intestinal juice, the swelling layer (c) begins to swell and increase in volume. At the same time, the pressure that prevails inside the pellets and acts on the layer providing delayed release increases. This pressure leads to the appearance of holes or bald spots in the sustained release layer, through which dissolved budesonide can diffuse from the inner layer (a) with the active substance to the outside. As the pellets continue to pass through the lower digestive tract, this process continues uninterrupted, ultimately ensuring complete release of budesonide from the pellets through the sustained release layer thus opened.

The release of the active substance begins only at a pH value above 6 with a short delay. The active substance is then released continuously and independently of the pH value for approximately 9 hours, at the end of which period budesonide is almost completely released from the pellets, which corresponds to an amount of more than 85% of the declared dose of the active substance. It is important that the release of the active substance does not occur abruptly or over a short period of time lasting from 1 to 2 hours, but more or less evenly over a period of approximately 8-10 hours, preferably 9 hours.

The release of budesonide as an active substance from the pellets according to the present invention is determined in the examples by performing in vitro release experiments using the following conditions and equipment:

testing equipment: apparatus with a paddle stirrer (apparatus 2 according to the European Pharmacopoeia);

- 3 043403 stirring speed: 75 rpm;

environments for conducting experiments:

(a) simulated gastric fluid (SGF):

Wednesday: 0.1 n. HCl, pH 1, 0.1% polysorbate 80;

volume: 900 ml;

experiment duration: 2 hours;

criterion: resistance to the action of gastric juice (lack of release);

(b) simulated intestinal fluid (SIF):

medium: phosphate buffer with 0.1% polysorbate 80 and an osmolarity of approximately 270 milliosmoles/kg and a pH of 6.5;

volume: 900 ml;

experiment duration: 16 hours;

criterion: complete release.

In separate experiments, the pellets were first kept in artificial gastric juice for two hours. Since in this case the pH value was set to 1.2, the essentially complete absence of release of the active substance, budesonide, indicates that the pellets were intact. After two hours, the pellets were removed from the artificial gastric juice and transferred into artificial intestinal juice with a pH value of 6.5. At the same time, the release of budesonide into the medium was analyzed. The pellets proposed in the invention are distinguished by the fact that during the first two hours (in artificial gastric juice) budesonide was practically not released. After two hours (in artificial intestinal juice), budesonide began to be released continuously, enabling the concept of colon targeting.

When testing compositions that are known from the prior art, the release of budesonide was observed partly already in the gastric juice. On the other hand, rapid release of budesonide was observed in the intestinal juice immediately in the first hours. This release profile is unsuitable for achieving the intended purpose, since budesonide is released primarily in the small intestine and possibly in the initial parts of the colon, but not primarily in the colon, as required.

For the production of pharmaceutical pellets, it is possible to use primarily two different methods described in the literature: coating of the original cores (production of multilayer pellets), as well as wet extrusion and melt extrusion. Multilayer pellets (according to the invention) are multi-coated starting pellets, the starting cores being, for example, active ingredient-containing sugar and starch pellets (so-called nonpareils). In this case, the active substance is first sprayed as a separate layer onto the surface of the nonpareils, and then subsequent functional layers are applied, which modify the release of the active substance. The application of individual layers or their application by spraying takes place in a continuous manner in a fluidized bed, as a result of which the pellets acquire their specific functionality.

According to the invention, multilayer pellets are used to implement the principle of targeting budesonide to the colon. The formation of the developed multilayer pellets occurs according to the following complex scheme:

Sugar and starch pellets as inert starting pellets with size/diameter 0.85-1.00 mm F

Layer 1: Application of budesonide to obtain pellets with active substance F

Layer 2: Application of a swelling layer F

Layer 3: Application of a layer providing delayed release of F

Layer 4: Application of a gastric juice-resistant layer to obtain gastric juice-resistant pellets F

Packaging in hard gelatin capsules or _________________stick bags_________________

When creating the invention, it was unexpectedly possible to establish that only a complex and systematic formation of the pellets proposed in the invention by sequentially forming the initial pellets, then the pellet with the active substance, then the pellet with a layer capable of swelling,

- 4 043403 then a slow-release pellet (pellet with a layer that provides a slow release) and finally a gastric resistant pellet allows for the delivery of budesonide to the colon, as well as its subsequent continuous release in it.

In addition to this strict structural formation of the multilayer pellets, another decisive feature of the solution proposed in the invention is the choice of suitable auxiliary substances for the individual layers. Only due to a certain consistency of structure and composition is it possible to optimally use the advantages of the multi-particulate form of administration with budesonide as the active substance, taking into account the implementation of the required principle of targeting the colon.

The individual layers of the preferred pellets according to the invention have, starting from the original cores, the following qualitative composition.

Pellets with active ingredient.

The active substance, i.e. budesonide is applied to the original pellets (sugar and starch pellets) in an aqueous-organic suspension consisting of water and isopropyl alcohol (mass ratio 80% by weight and 20% by weight), as well as other components, which are lactose monohydrate ( function: filler), polyvinylpyrrolidone (PVP) type Kollidon® K25 (function: binder), polyoxyethylene(20)-sorbitan monooleate (Polysorbate® 80, Tween 80) (function: wetting agent) and talc (function: release agent). The proportion of the solid phase in the suspension is about 27% by weight. After budesonide is applied and the pellets are dried, the budesonide is bonded to the original pellets.

Pellets with a layer capable of swelling.

The active ingredient pellets are then sprayed with an alcohol suspension of homopolymeric cross-linked polyacrylic acid type A (carbomer, viscosity [0.5%]: 4000 to 11000 mPa-s, approximate relative molecular weight: 1250000) (function: swelling agent), polyvinylpyrrolidone (PVP) type Kollidon® K25 (function: binder) and talc (function: anti-adhesive). In this way, a swelling layer is applied. Isopropyl alcohol is used as a solvent. The proportion of the solid phase in the suspension is about 11% by weight.

Pellets with a layer that provides a slow release.

At the next stage, a film coating is applied to the pellets with a combination of polymers that modifies the release of the active substance. In this case, a combination of ammonium methacrylate copolymer type B (copolymer of methyl, ethyl and 2-trimethylaminoethyl esters of (meth)acrylic acid with a ratio of ethyl acrylate, methyl methacrylate and trimethylammonioethyl methacrylate 1: 2: 0.1, Eudragit® RS 12.5) and copolymer ammonium methacrylate type A (copolymer of methyl, ethyl and 2-trimethylaminoethyl esters of (meth)acrylic acid with a ratio of ethyl acrylate, methyl methacrylate and trimethylammonioethyl methacrylate 1:2:0.2, Eudragit® RL 12.5). Both polymers form water-insoluble but permeable films and are added to a spray solution, respectively a spray suspension in a 12.5% by weight solution in isopropyl alcohol (60% by weight) and acetone (40% by weight) with sodium lauryl sulfate in as a wetting agent. The solution to be sprayed or the suspension to be sprayed additionally also contains triethyl citrate (function: plasticizer) and talc (function: release agent). The proportion of the solid phase in the aqueous-organic spray solution, or the aqueous-organic spray suspension, is about 14% by weight. The components of the sustained release layer are suspended in this spray solution, respectively in this spray suspension, which consists of isopropyl alcohol (about 88% by weight) and water (about 12% by weight). After film coating and drying of the pellets, the sustained release layer is placed directly on the swellable layer. Gastric acid-resistant pellets At the last stage, a gastric juice-resistant coating is applied to the pellets. To do this, use an aqueous-organic solution of Eudragit® L (a copolymer of polymethacrylic acid and polymethyl methacrylate with a 1:1 ratio between polymethacrylic acid and polymethyl methacrylate). This polymer forms a coating soluble in intestinal juice. In addition to the anionic polymer, the spray solution or spray suspension also contains triethyl citrate (function: plasticizer) and talc (function: release agent), whereby the solid phase content is approximately 15%. The solvent consists of 85% by weight isopropyl alcohol and 15% by weight of water. Applying this layer to the pellets reliably prevents the release of the active substance in the stomach.

The coating of pellets in individual layers is preferably carried out in fluidized bed plants. Since the individual layers are applied from the material in the form of liquids or suspensions, the liquid phase used as a carrier must be removed. Removal of the liquid phase occurs through reasonable ventilation, and the ambient temperature can be raised moderately, but only to a temperature at which no undesirable side reactions occur.

The required functionality of multilayer pellets is realized as a result of optimal, sequential dissolution and swelling of the applied layers along the digestive tract: immediately along

- 5 043403 When the pellets reach the small intestine, the polymer of the outer layer (e) first dissolves. Intestinal digestive juices penetrate through the permeable sustained release layer (d) into the pellets. The prevailing pH conditions in the small intestine (pH less than 5.5 to about 7.2) cause the polyacrylic acid to swell, i.e. polymer layer capable of swelling. The associated increase in volume is accompanied by an increase in internal pressure on the sustained release layer, which results in continuous bald spots. In this way, holes are formed through which budesonide, dissolved in the digestive juice, can diffuse out. In this way, the active substance dissolved in the digestive juice can diffuse outward over a long period of time, thereby being released at the site of its action and exerting a local effect. This process occurs depending on time and begins in full when the pellets reach the colon.

Multilayer pellets formed in this way can be packaged in hard gelatin capsules in the appropriate dosage. The qualitative and quantitative composition of one such preferred 9 mg budesonide capsule is summarized in Table 1. 1. The solvents used are not a component of the composition, since they are removed by drying in the form of volatile components during the process.

When developing multilayer pellets, various prototypes were made according to the preferred option, which had basically one combination of components, as is presented in more detail in Table. 1.

Table 1

Composition of the preferred multilayer pellets containing budesonide in capsule according to the invention | Composition [mg] |

Component Function Amount per capsule 1. Pellets with active ingredient Budesonide medically active ingredient 9.00 mg Lactose monohydrate filler 36.00 mg Polyvinylpyrrolidone (Kollidon K25) binder 2.70 mg Fully xethylene(20)-sorbitan monooleate (polysorbate 80; tween 80) wetting agent 0.71 mg Talc release agent 16.50 mg Sugar and starch pellets original pellets 260.00 mg Total: 324.91 mg 2. Pellets with a swelling layer Carbomer swelling agent 24.00 mg (TO) Polyvinylpyrrolidone (Kollidon K25) binder 18.00 mg Talc release agent 18.00 mg Total: 384.91 mg 3. Pellets with a layer that provides a slow release Eudragit® RS 12.5 sustained release polymer 6.30 mg Eudragit® RL 12.5 sustained release polymer 2.70 mg Triethyl citrate plasticizer 0.90 mg Talc release agent 7.20 mg Total: 402.01 mg 4. Gastric juice-resistant pellets Eudragit® L enteric juice soluble polymer 16.00 mg Triethyl citrate plasticizer 1.60 mg Talc anti-caking agent 8.00 mg Total: 428.81 mg 5. Hard gelatin capsule Gelatin with titanium dioxide capsule shell 97.00 mg Total: 525.81 mg

The in vitro release profile of the active substance from the pellets proposed in the invention is demonstrated in example 1. Such pellets have the required criteria for resistance to the action of jelly

- 6 043403 milk juice and continuous release in artificial intestinal juice respectively satisfy the specified parameters. Example 2 demonstrates that the desired release of budesonide from the pellets can surprisingly be achieved solely by the combined use of swelling and sustained release layers. Only through the sequential application of swelling and sustained-release layers onto the active ingredient pellets as described for the solution according to the invention is it possible to deliver budesonide to its site of action in the colon.

Negative example (extruded pellet, example 4).

In this case, an attempt was made to develop a pharmaceutical composition with the required release profile of the active substance according to the technological option by extrusion followed by spheronization.

Extruded pellets (a negative example) mean powder agglomerates, with the particles being obtained by wet extrusion of a powder mass or its extrusion from a melt followed by spheronization. Extruded pellets were used only as a control. Extruded pellets usually allow them to be supplied with a higher dosage of the active substance, which, however, is not essential according to the present invention, since topical corticoids are used in a low dosage. For extrusion, screw extruders are used, in which, by applying pressure to a wet or molten mass, it is forced through holes of a certain size. The extrudates obtained in this way are cut into cylindrical pellets and then rounded into balls. These pellets can also be coated in fluid bed plants. Unlike multilayer pellets, extruded pellets are produced in a periodic manner. Important factors for choosing the type of pellet are, among other things, the physicochemical properties of the active substance being processed, its required dosage and the required mechanism for its release in order to obtain the target release profile. According to the invention, extruded pellets are not used.

The formation of extruded pellets was carried out in accordance with the following scheme regarding the quality composition and manufacturing:

Granulate with active substance g Extrusion f Spheronization F

Gastric juice-resistant pellets F

Packaging in hard gelatin capsules or _________________stick bags_________________

Granules with active substance.

A powder mixture of budesonide, lactose monohydrate (function: filler), carrageen (Gelcarin GP911 NF) (function: matrix former and extrusion aid) and calcium chloride (function: enhanced gelation or matrix formation) was wet granulated using an aqueous granulator. liquids. The granulating liquid also contained polyvinyl acetate (Kollicoat SR 30D) (function: slow release polymer and matrix former). After sifting the wet mass, extrusion was carried out.

Extrusion and spheronization.

The wet mass was pressed in a screw extruder at 65 °C through a grid (a matrix with certain holes with a diameter of 1.0 to 2.2 mm) and then cut into cylindrical molded blanks. These molded blanks were rounded at a maximum temperature of 50°C, dried and finally the fine fraction (mesh size 1000 µm) was separated by sieving.

Gastric acid resistant pellets.

At the last stage, a coating resistant to gastric juice is applied to the round blanks. To do this, use an aqueous suspension of a mixture of Eudragit® FS 30 D (copolymer of methacrylic acid and methyl (meth)acrylate) and Eudragit® L 30 D 55 (copolymer of methacrylic acid and ethyl acrylate) in a ratio of approximately 80% by weight and 20% by weight, respectively. . Both polymers form an intestinal juice-soluble coating. In addition to these components, the aqueous suspension also contains triethyl citrate (function: plasticizer), glycerol monostearate (function: anti-caking agent) and polyoxyethylene (20) sorbitan monooleate (Polysorbate® 80; Tween 80) as a wetting agent, as well as sodium lauryl sulfate, according to with which the share of the solid phase is about 21%. Applying this layer to extruded pellets reliably prevents the release of the active substance in the stomach.

The extruded pellets formed in this way can be packaged in hard gelatin capsules in the appropriate dosage. Qualitative and quantitative composition of one such cap

- 7 043403 sula with a dose of budesonide 9 mg is summarized in table. 2. The solvents used are not a component of the composition, since they are removed by drying in the form of volatile components during the process.

The purpose of developing such a composition was to ensure swelling of the pellet matrix after reaching the small intestine and after the dissolution of the outer layer polymers by the digestive juices that penetrated inside. In this way, gradual erosion of the particles should occur as they pass through the intestinal sections, thereby leading to a continuous release of the active substance.

The in vitro release profile of such extruded pellets is demonstrated in Example 4. Such extruded pellets do not meet the required criteria for targeted topical administration to the colon. Thus, this approach to the development of a pharmaceutical composition does not lead to the implementation of the required principle of targeting the colon.

table 2

Composition of a capsule containing extruded pellets with budesonide (negative example)

Composition [mg] Component Function Amount per capsule 1. Granules with active ingredient Budesonide medically active ingredient 9.00 mg Polyvinyl acetate (Kollicoat SR30D) sustained release polymer, matrix former 52.50 mg Lactose monohydrate binder 28.50 mg Carrageen (Gelcarin GP911 NF) matrix former, extrusion aid 240.00 mg Calcium chloride gelling and matrix former 20.00 mg Total: 350.00 mg 2. Gastric juice-resistant pellets Eudragit® FS 30 D enteric juice soluble polymer 48.00 mg Eudragit® L 30 D 55 enteric juice soluble polymer 12.00 mg Triethyl citrate (Citrofol AL) plasticizer 2.80 mg Glycerol monostearate anti-caking agent 3.10 mg Polyoxyethylene(20)sorbitan monooleate (Polysorbat® 80; Tween 80) wetting agent 1.10 mg Total: 417.00 mg 3. Hard gelatin capsule Gelatin with titanium dioxide capsule shell 97.00 mg Total: 514.00 mg

The dissolution process of the composition according to the invention is schematically illustrated in FIG. 3.

In contrast to the negative comparative example, when using the multilayer pellets proposed in the invention, a completely different picture was observed with the level of the active substance in the blood. After dissolution of the gastric juice-resistant film, in this case the complex mechanism already described above for releasing the active substance is triggered. Plasma levels of budesonide do not increase sharply, but exhibit a modified, slow and prolonged pattern of change over several hours.

The results of pharmacokinetic studies confirm the results of in vitro studies in example 1 and clearly demonstrate the fact that the active substance is released from multilayer pellets throughout the entire passage through the colon and therefore reaches those places where it can exert its effect on the colon mucosa .

In an open, single-center, randomized pharmacokinetic study, the multilayer pellets proposed in the invention were compared with Budenofalk hard capsules with a dosage of the active substance of 3 mg, known from the prior art. Average plasma concentrations of the active substance were measured over time after a single oral dose of the active substance at a dose of 9 mg, while the bioavailability of budesonide from the control composition is known. The results of a comparative pharmacokinetic study are presented in example 3.

- 8 043403

As follows from the data on plasma levels of the active substance in example 3, the composition described in the prior art is unsuitable for targeted delivery of budesonide to the colon. The level of budesonide in the blood increases extremely quickly and significantly. This type of change in the level of budesonide in the blood is undesirable for the implementation of the principle of targeting the large intestine, since the active substance does not reach the site of its action in sufficient concentration, and is already largely absorbed in the upper parts of the small intestine and is therefore no longer available for local, respectively local action in the colon. This behavior in vivo can be explained by the composition of this composition. The Budenofalk hard capsules with a dosage of 3 mg of active substance described in the prior art contain only pellets with budesonide coated with a gastric juice-resistant coating. After passing through the stomach, this coating dissolves with some delay, after which the active substance is released immediately and completely. As a result, the active substance does not reach the colon. The results of a clinical study testing this prior art composition also indicate that Budenofalk 3 mg hard capsules (described in WO 95/08323) are not suitable for the treatment of ulcerative colitis. However, it has not been able to demonstrate efficacy at least comparable to mesalazine therapy in the treatment of patients with mild to moderate ulcerative colitis (Gross et al., 2011).

The effectiveness and tolerability of the new multilayer pellets were also tested in an open clinical study. As described in example 5, the results obtained clearly confirm the clinical effectiveness of the pellets. The results obtained demonstrate that the pellets proposed in the invention deliver the active substance to the required destination and allow successful treatment of patients who suffer from ulcerative colitis and whose treatment with mesalazine failed to achieve satisfactory results (immunity to mesalazine).

In contrast to multilayer pellets, the pattern of changes in the level of active substance in the blood revealed during the pharmacokinetic study in example 4 when using extruded pellets (negative example) is more similar to the state of the art than to the required target profile for the composition releasing the active substance in the colon. Levels of budesonide in the blood increase even faster than with the control composition, and accordingly also quickly decrease again, whereby the release of the active substance is completed before reaching the colon, and budesonide, apparently, does not reach it in any significant way. quantities.

Important aspects of the present invention are illustrated in the accompanying drawings and are explained below with reference thereto.

In fig. Figure 1 schematically shows the digestive tract.

In fig. 2 schematically shows the structure of the pellets according to the invention for sustained release of the active substance.

In fig. 3 illustrates the principle of operation of the pellets proposed in the invention, depending on a specific area in the digestive tract.

In fig. 4 graphically presents the in vitro release profiles of the active substance from different pellets, comparing pellets with the active substance, pellets with a swelling layer, pellets with a layer providing a slow release, and pellets resistant to the action of gastric juice regarding release from of the active substance, and the composition proposed in the invention is designated as pellets resistant to the action of gastric juice. The experimental conditions are explained in more detail in Example 1. The abbreviation SGF stands for artificial gastric juice and the abbreviation SIF stands for artificial intestinal juice.

In fig. 5 graphically shows the in vitro release profile of the active substance from pellets according to the invention with and without a swelling layer. The experimental conditions are explained in more detail in example 2. Shown in Fig. 5, the experimental results confirm that a sustained release layer without an underlying swelling layer does not provide the ability to release the active substance.

In fig. Figure 6 shows the in vivo release profile obtained from open-label, randomized pharmacokinetic studies. We are talking about data obtained in vivo, which explains possible discrepancies with data obtained in vitro.

In fig. 7 is a diagram which confirms that effective treatment of the entire colon is in fact possible with the compositions of the invention. In example 6, the effectiveness and tolerability of the budesonide pellets proposed in the invention were examined, while in FIG. Table 7 shows the proportion of patients with clinical remission depending on the location of the specific focus of the disease.

In fig. 8 graphically illustrates the effectiveness of the compositions of the invention as measured by the number of bloody stools per week. Shown in Fig. 8 results confirm that the treatment effectively reduced the number of bloody stools per week.

- 9 043403

Using the materials and methods described above, various in vivo pharmacokinetic profiles were determined. The following examples illustrate the invention.

Example 1. Experiments with the release of budesonide in vitro from multilayer pellets containing it (the composition proposed in the invention).

In order to demonstrate the effect, as well as the influence of individual layers on the release of the active ingredient, in vitro experiments were carried out with active ingredient pellets, swellable layer pellets, sustained release layer pellets and gastric resistant pellets from the same batch. The study was carried out with a quantity of pellets that corresponds to a dose of budesonide of 9 mg. The outer hard gelatin capsule has no effect on the release of the active substance from the pellets and was therefore not included in these experiments.

Experiments with the release of the active substance in vitro from pellets covered a two-stage study. At the first stage, the pellets were studied in artificial gastric juice (pH 1.2) for two hours, followed by the transfer of test samples into artificial intestinal juice (pH 6.5), as well as further study of the release of the active substance for 7 hours. provide conditions for immersing the test samples in both media; they contained 0.1% polysorbate 80 as a wetting agent. The parameters for the study of the release of the active substance were as follows:

release test apparatus: apparatus 2 according to the European Pharmacopoeia, described in chapter 2.9.3 (paddle method);

release medium volume: 900 ml artificial gastric juice (SGF) and 900 ml artificial intestinal juice (SIF);

rotation speed: 75 rpm; temperature: 37.0±0.5°C;

test environment:

artificial gastric juice (SGF): 0.1 N. HCl with 0.1% polysorbate 80, pH 1.2, artificial intestinal fluid (SIF): phosphate buffer (KH ₂ PO ₄ , NaCl, NaOH) with 0.1% polysorbate 80, pH 6.5, batch of test samples: 180013457;

number of tested samples: N=6;

sampling:

pellets with active ingredient: 5, 10, 15, 30, 45, 60 and 120 min in artificial gastric juice, pellets with a swelling layer: 5, 10, 15, 30, 45, 60 and 120 min in artificial gastric juice, as well as 15, 30, 45, 60, 90 and 180 minutes in artificial intestinal juice, pellets with a layer that provides sustained release: 15, 30, 45, 60, 90 and 120 minutes in artificial gastric juice, 30, 90, 150, 210, 270, 330, 390 and 420 min in artificial intestinal juice, resistant to the action of gastric juice, pellets: 15, 30, 45, 60, 90 and 120 min in artificial intestinal juice, 30, 90, 150, 210, 270, 330, 390 and 420 min in artificial intestinal juice;

determination of budesonide content: HPLC/UV (λ=255 nm).

In fig. Figure 4 graphically shows the in vitro release profiles of the active substance from individual pellets. Pellets with the active substance (uncoated) already completely release the active substance in the gastric juice. After 5 minutes, more than 85% of budesonide is dissolved in it. Pellets with a swelling layer exhibit a slight release of the active substance during a two-hour test in gastric juice (less than 5%) and, after transfer to the intestinal juice, completely release the active substance within 15 minutes (at least 85%). This behavior of these pellets reflects the physicochemical properties of the polyacrylic acid (carbomer) used, which, due to its pKs value of about 6, is either insoluble, swelling, or soluble in it, depending on the pH value of the medium. Under conditions of artificial gastric juice (pH 1.2), the high molecular weight polymer of acrylic acid is presented in a protonated form and is insoluble. The swelling layer remains on the pellets. After transfer to a buffer simulating intestinal juice, the pH value increases to a value in the pKs range and leads to partial deprotonation of carboxylic acids. As a result, water molecules can be incorporated into the narrow polymer framework, which leads to the formation of a gel framework and thereby swelling. However, in the absence of a sustained release layer, this effect of the swelling layer does not occur and budesonide is therefore released extremely quickly.

Only after applying the sustained release layer to pellets with a swelling layer can the pellets fully exhibit the effect described above (see FIG. 4: release profile of the active ingredient from pellets with a sustained release layer). The release profile of the active substance from pellets with a layer providing sustained release in intestinal juice clearly changes and becomes the required S-shape

- 10 043403 form, which is associated with the delayed release of budesonide. After approximately 120 minutes in the gastric juice, complete release of the active substance from such pellets with a sustained release layer is then achieved in the intestinal juice after a further approximately 150 minutes. Thus, there is no premature release of the active substance in the gastric juice.

However, only the final composition with the additional gastric acid-resistant coating ultimately exhibits the release profile of the active substance corresponding to the composition proposed in the invention required for the implementation of the principle of targeting the colon. Release of budesonide begins approximately 90 minutes after passage of the stomach. Thus, there is no premature release of the active substance in it. After this time, the pellets have passed through the small intestine and reached the large intestine. After this, the active substance is continuously released with sigmoidal kinetics for at least seven hours. In general, the release of the active substance from the composition proposed in the invention in such an in vitro system occurs in accordance with the following criteria:

SGF (simulated gastric fluid, i.e. artificial gastric juice), pH 1.2:

after 2 hours: no release, which corresponds to resistance to gastric juice;

SIF (simulated intestinal fluid, i.e. artificial intestinal juice), pH 6.5:

after 270 minutes: release 10-30%, after 330 minutes: release 40-70%, after 540 minutes: release at least 80%.

The results of HPLC/UV determinations of the active ingredient content of active ingredient pellets, swellable layer pellets, sustained release layer pellets and gastric acid resistant pellets confirm that the pellets tested in separate in vitro release studies of the active substance contained budesonide in the required amount of 9 mg (corresponding to its content in one capsule).

Pellets with active ingredient: 100.0%.

Pellets with a swelling layer: 101.9%.

Slow release layer pellets: 101.6%.

Gastric acid resistant pellets: 100.3%.

Example 2 In vitro release experiments of budesonide from sustained release layer pellets with or without a swelling layer.

In order to demonstrate the interaction of the swelling layer and the sustained release layer in the release of the active substance according to the present invention, budesonide containing pellets with a sustained release layer, with or without a swelling layer, were prepared according to the following composition.

Composition [mg] Component Amount per dose of budesonide 9 mg Batch R020 Batch R029 1. Pellets with active ingredient Budesonide 9.00 mg 9.00 mg Lactose monohydrate 36.00 mg 36.00 mg Polyvinylpyrrolidone (Kollidon K25) 2.70 mg 2.70 mg Polyoxyethylene(20)-sorbitan monooleate (Polysorbate 80, Tween 80) 0.71 mg 0.71 mg Talc 16.50 mg 16.50 mg Sugar and starch pellets 260.00 mg 260.00 mg Total: 324.91 mg 324.91 mg

- 11 043403

Composition [mg] Component Amount per dose of budesonide 9 mg Batch R020 Batch R029 2. Pellets with a swelling layer Carbomer — 24.00 mg Polyvinylpyrrolidone (Kollidon K25) — 18.00 mg Talc — 18.00 mg Total: 324.91 mg 384.91 mg 3. Pellets with a layer that provides a slow release Eudragit® RS 12.5 10.50 mg 10.50 mg Eudragit® RL 12.5 4.50 mg 4.50 mg Triethyl citrate 1.50 mg 1.50 mg Talc 12.0 mg 12.0 mg Total: 353.41 mg 413.41 mg

In terms of the composition of the layer with the active substance and the swelling layer, batch R029 corresponds to the invention. However, in comparison with the invention, the application of the sustained release layer was intentionally increased by 1.7 times. Using this test batch as an example, it must be demonstrated that, with regard to the release of the active substance, both of these layers function in their interaction, respectively in their joint action, even under extreme conditions and that there is therefore a reliable effect of the swelling layer. The pellets of batch R020 do not contain a layer capable of swelling, and this batch served as a control. With the exception of this difference, the pellets of the control and test batches had identical composition. The pellets of both batches were not covered with a film resistant to gastric juice.

The parameters for the study of the release of the active substance were as follows:

release test apparatus: apparatus 2 according to the European Pharmacopoeia, described in chapter 2.9.3 (paddle method);

volume of release medium: 900 ml artificial intestinal juice (i.e., resistance to gastric juice was not tested);

rotation speed: 75 rpm; temperature: 37.0±0.5°C;

test environment:

artificial intestinal juice (SIF): phosphate buffer (KH ₂ PO ₄ , NaCl, NaOH) with 0.1% polysorbate, pH 6.5;

batches of test samples:

test lot: R029, control lot: R020;

number of tested samples: N=2;

sampling and determination of budesonide content: continuously by UV measurement directly in the process (online) (λ=255 nm).

In fig. Figure 5 graphically shows the in vitro release profiles of the active substance from the pellets of the test and control batches. The effect of the swelling layer is impressive and completely unexpected. While no budesonide was released from the control batch pellets over the course of hours, its release profile from the test batch pellets was consistent with that of the invention. Thus, the simultaneous presence of the swelling and sustained release layers allows for the required sustained release of budesonide from the pellets. The sustained release layer alone is not effective in this regard. Thus, only the combination of the swelling and sustained-release layers proposed in the invention is capable of delivering budesonide to the site of its action, and therefore both of these layers should be considered together as the release-controlling components of the composition.

Example 3. Pharmacokinetic profiles of a budesonide-containing multilayer pellet capsule (composition of the invention) and a budesonide-containing capsule from the prior art/control in in vivo experiments.

During the first phase of an open-label, randomized study of the pharmacokinetics of the composition proposed in the invention, 12 healthy male subjects took a single dose of the composition proposed in the invention on an empty stomach (symbol BUX-PVII; 9 mg budesonide) and budesonide-containing capsules from the prior art (3x3 mg budesonide per one dose as control pellets). Important pharmacokinetic parameters are presented in the table below. 3.

- 12 043403

Table 3

Application of the pellets according to the invention (BUX-PVII) Use of control pellets (3x3 mg budesonide) Stach (ng/ml) tmax (h) Stach (ng/ml) tmax (h) N 12 12 12 12 Mean (SD) 0.928 (0.755) 7.75 (3.71) 4.60 (4.85) 5.26 (0.81) Min 0.193 2.00 1.04 4.50 Max 2.62 13.00 16.1 6.50

In table 3 N means the number of subjects, Mean (SD) means the average value/standard deviation, Min means the minimum value, Max means the maximum value. In table 3 for each of the values of Cmax and tmax the corresponding average value of the maximum level of budesonide in the blood plasma of individual patients is indicated, regardless of the moment of measurement.

The results of the in vivo pharmacokinetic study are presented graphically in FIG. 6. Compared with the values indicated in table. 3, in fig. 6 individual values were measured at a specific point in time and the corresponding average was calculated. Therefore, the values indicated in table. 3 are different from the values in FIG. 6.

Example 4. Pharmacokinetic profiles of a capsule containing budesonide extruded pellets (negative example) in in vivo experiments.

In a phase 1, open-label, randomized study of the pharmacokinetics of budesonide-containing extruded pellets, 16 healthy male subjects administered a single dose of budesonide-containing extruded pellets (with a dosage of 9 mg budesonide per gelatin capsule) in a single, fasted state. Important pharmacokinetic parameters are presented in the table below. 4. In table. 4 for each of the values of C _max and t _max the corresponding average value of the maximum level of budesonide in the blood plasma of individual patients is also indicated, regardless of the moment of measurement.

Table 4

Application of budesonide-containing extruded pellets (BUX-E) Stach (ng/ml) tmax (h) N 16 16 Mean (SD) 2.478 (0.9683) 3.97 (1.407) Min 0.913 2.00 Max 3.95 7.00

Given in table. 3 and 4, the data confirm that only when using the pellets proposed in the invention, it was possible to achieve a late release of the active substance (t _max = 7.75), whereas when using pellets from the prior art, a t _max value of 5.26 was observed, and when using extruded pellets - tmax value equal to 3.97.

Example 5.

The results obtained in this example are presented graphically in Fig. 6. In FIG. Figure 6 shows a graph of changes in plasma budesonide levels as a function of time based on the results of open-label, single-center, randomized pharmacokinetic studies. The graph shows the corresponding average values of budesonide plasma levels of all patients at the corresponding measurement time. In contrast, for those indicated in table. 3, respectively 4 values do not matter when the corresponding Cmax, respectively tmax value was measured in a particular patient. Along with the composition proposed in the invention, other prototypes of a medicinal composition containing budesonide were tested. Schematically shown in Fig. 2, the composition with the symbol BUX-PVII allows for optimal implementation of the principle of targeting the colon. The achieved in vivo release profile of the active substance should, after a delay phase, ensure its release evenly over a longer period of time, while at the same time low levels in serum and plasma. Mean plasma concentrations of budesonide were measured after a single dose of budesonide-containing pellets over time. In fig. 6 graphically presents the results for two control compositions (BUX-E and control) and for the composition proposed in the invention (BUX-PVII).

Only the composition according to the invention exhibits the required release profile of the active substance: a clear delay in the onset of its release (about 3 hours), a plateau-like plasma level of budesonide over a long period of time (for about 8 hours) and, in comparison, a clearly lower level of budesonide in plasma .

In this study, the plasma concentration profiles of budesonide when using the drug composition proposed in the invention were more homogeneous over time, with little variation between individual participants in the clinical study.

Example 6.

An 8-week open-label clinical trial tested the efficacy and

-

Claims (3)

tolerability of a prototype of a new oral formulation of 9 mg budesonide given once daily. The goal of this study was to achieve clinical remission, defined by a disease activity score of no more than 4 (Colitis Activity Index, CAI) and by subscores 1 and 2 of the ACI, each of which is equal to 0. A subscore of ACI with a value of 0 describes a stool frequency of less than 18 per day. week, and IAC sub-indicator 2 with a value of 0 describes a condition in which there is no or a maximum of 1 bloody stool per week. This study treated patients with active ulcerative colitis who had failed treatment with a mesalazine-containing drug as standard therapy. Such pretreatment must have been discontinued prior to the start of the clinical trial and prior to the first administration of the study drug. A total of 61 patients were treated, of whom 52 patients completed the study as per protocol. Proportion of patients who achieved the study goal, i.e. primary (hard) endpoint is given in table. 5. The so-called FAS group combines all patients who were treated with the test drug at least once. The so-called RR group includes those patients who completed the study according to the protocol. Both groups analyzed achieved approximately 50% remission rates in this mesalazine-refractory study population. Table 5 Primary endpoint analysis Analyzed Number (%) of patients with clinical remission of the 1st group _______________type at the end of the study______________ ____Total_____ CI ¹ 95% by the Clopper-Pearson method N η % F.A.S. 61 29 47.5 [34.60%, 60.73%] RR 52 28 53.8 [39.47%, 67.77%] Note. CI means confidence interval; FAS (full analysis set) means full analysis population; PP (from the English per protocol set) means a sample of patients whose data corresponds to the protocol. In this study, it was possible to demonstrate that the composition of the invention actually has an effective effect throughout the entire colon (Fig. 7). Concordant efficacy, as measured by clinical remission rates, did not show any significant variation across colonic regions. It was also possible to effectively reduce the number of stools per week and the number of bloody stools per week (Fig. 8). In this study, it was possible to effectively and safely use the composition proposed in the invention. Safety of use is confirmed by measuring cortisol levels in the morning. The measured mean values remained within the normal acceptable range of 6.2 to 18 μg/dL. CLAIM 1. Multilayer coated pellet for sustained release of the active substance into the distal colon, having the following components: a) the original pellet, which consists only of inert material and does not contain any pharmaceutically active active substance inside; b) a layer with the active substance, which is applied directly to the original pellet a) and, in addition to the active substance, contains film-forming auxiliary substances; c) a swelling layer, which is applied directly to layer b) with the active substance and contains swelling materials that swell upon contact with intestinal juice; d) a sustained release layer which does not dissolve on contact with intestinal juices but becomes permeable to liquids and which is applied directly to layer c); e) an outer coating that is insoluble at a pH value less than 5.5, but readily soluble at a pH value greater than 6.0, and is applied directly to layer d), wherein the active substance is budesonide or a pharmaceutically acceptable salt thereof. 2. Pellets according to claim 1, characterized in that the initial pellet (a) has an average diameter from 0.2 to 2.0 mm, with at least 90% of the particles having dimensions within the specified limits, and the initial pellets have a spherical shape with a homogeneous surface. 3. Pellets according to claim 2, wherein at least 95% of the particles have sizes that range from 0.2 to 2.0 mm. -